Comparison of the structural organisation of reeler hippocampal CA1 region with wild type CA1.

The dendritic pattern defines the input capacity of a neuron. Existing methods such as Golgi impregnation or intracellular staining only label a small number of neurons. By using high-resolution imaging and 3D reconstruction of green fluorescent protein-expressing neurons, the present study provides an approach to investigate the anatomical organization of dendritic structures in defined brain regions. We characterized the structural organization of dendrites in the CA1 region of the mouse hippocampus by analyzing Sholl intersections, dendritic branches, branching and orientation angles of dendrites, and the different types of spine on the dendritic branches. Utilizing this quantitative imaging approach, we show that there are differences in the number of Sholl intersections and in the orientation of apical and basal dendrites of CA1 pyramidal neurons. Performing 3D reconstructions of the CA1 region of the reeler hippocampus, we show that neurons of this mutant display an arbitrary orientation of apical dendrites at angles ranging from -180 to +180 degrees in contrast to wild-type mice that show a preferred orientation angle. This methodology provides a way of analyzing network organization in wild type and mutant brains using quantitative imaging techniques. Here, we have provided evidence that in reeler a sparse, weakly connected network results from the altered lamination of CA1 pyramidal neurons and the variable orientation of their dendrites.

Gonadal Hormones Rapidly Enhance Spatial Memory and Increase Hippocampal Spine Density in Male Rats

17β-estradiol (E2) rapidly, within minutes, activates behaviors and cognition by binding to membrane estrogen receptors, activating cell signaling cascades and increasing dendritic spines. In female rodents, E2 enhances spatial memory within 2–4 hours, and spine density is increased in the CA1 area of the hippocampus within 30–60 minutes. Although chronic gonadal hormone treatments in male rats alter cognition and spines/spine synapses and acute hormone effects occur in hippocampal slices, effects of acute, in vivo hormone administration in males are unknown. Therefore, we assessed rapid effects of E2 (20 μg/kg) and testosterone (T) (750 μg/kg) on spatial memory using the object placement task and on hippocampal spine density using Golgi impregnation. Orchidectomized rats received hormones immediately after the training trial and were tested for retention 2 hours later. Vehicle-injected orchidectomized males spent equal time exploring objects in the old and new locations, but E2- or T-treated subjects spent more time exploring objects at the new location, suggesting enhanced memory. Both hormones also increased spine density in CA1, but not the dentate gyrus, by 20%–40% at 30 minutes and 2 hours after injections. This report is the first, to our knowledge, to show E2 and T enhancements of memory and spine density within such a short time frame in male rats.

Golgi Analysis of Neuron Morphology in the Presumptive Somatosensory Cortex and Visual Cortex of the Florida Manatee (Trichechus manatus latirostris)

The current study investigates neuron morphology in presumptive primary somatosensory (S1) and primary visual (V1) cortices of the Florida manatee (Trichechus manatus latirostris) as revealed by Golgi impregnation. Sirenians, including manatees, have an aquatic lifestyle, a large body size, and a relatively large lissencephalic brain. The present study examines neuron morphology in 3 cortical areas: in S1, dorsolateral cortex area 1 (DL1) and cluster cortex area 2 (CL2) and in V1, dorsolateral cortex area 4 (DL4). Neurons exhibited a variety of morphological types, with pyramidal neurons being the most common. The large variety of neuron types present in the manatee cortex was comparable to that seen in other eutherian mammals, except for rodents and primates, where pyramid-shaped neurons predominate. A comparison between pyramidal neurons in S1 and V1 indicated relatively greater dendritic branching in S1. Across all 3 areas, the dendritic arborization pattern of pyramidal neurons was also similar to that observed previously in the afrotherian rock hyrax, cetartiodactyls, opossums, and echidnas but did not resemble the widely bifurcated dendrites seen in the large-brained African elephant. Despite adaptations for an aquatic environment, manatees did not share specific neuron types such as tritufted and star-like neurons that have been found in cetaceans. Manatees exhibit an evolutionarily primitive pattern of cortical neuron morphology shared with most other mammals and do not appear to have neuronal specializations for an aquatic niche.

Morphology of neurons of human subiculum proper

Subiculum proper is an archicortical structure of the subicular complex and presents the place of origin of great majority of axons of the whole hippocampal formation. In contrast to the hippocampus which has been intensively studied, the data about human subiculum proper are quite scarce. The aim of our study was to indentify morphological characteristics of neurons of the human subiculum proper. The study was performed on 10 brains of both genders by using Golgi impregnation and Nissl staining. The subiculum has three layers: molecular, pyramidal and polymorphic layer. The dominant cell type in the pyramidal layer was the pyramidal neurons, which had pyramidal shaped soma, multiple basal dendrites and one apical dendrite. The nonpyramidal cells were scattered among the pyramidal cells of the pyramidal layer. The nonpyramidal cells were classified on: multipolar, bipolar and neurons with triangular-shaped soma. The neurons of the molecular layer of the human subiculum were divided into groups: bipolar and multipolar neurons. The most numerous cells of the polymorphic layer were bipolar and multipolar neurons.

The Spine Loss Paradox: Clues to Mechanisms and Meaning

A Cellular Mechanism for Dendritic Spine Loss in the Pilocarpine Model of Status Epilepticus. Kurz JE, Moore BJ, Henderson SC, Campbell JN, Churn SB. Epilepsia 2008 May 8. [Epub ahead of print] PURPOSE: Previous studies have documented a synaptic translocation of calcineurin (CaN) and increased CaN activity following status epilepticus (SE); however, the cellular effect of these changes in CaN in the pathology of SE remains to be elucidated. This study examined a CaN-dependent modification of the dendritic cytoskeleton. CaN has been shown to induce dephosphorylation of cofilin, an actin depolymerization factor. The ensuing actin depolymerization can lead to a number of physiological changes that are of interest in SE. METHODS: SE was induced by pilocarpine injection, and seizure activity was monitored by video-EEG. Subcellular fractions were isolated by differential centrifugation. CaN activity was assayed using a paranitrophenol phosphate (pNPP) assay protocol. Cofilin phosphorylation was assessed using phosphocofilin-specific antibodies. Cofilin–actin binding was determined by coimmunoprecipitation, and actin polymerization was measured using a triton-solubilization protocol. Spines were visualized using a single-section rapid Golgi impregnation procedure. RESULTS: The immunoreactivity of phosphocofilin decreased significantly in hippocampal and cortical synaptosomal samples after SE. SE-induced cofilin dephosphorylation could be partially blocked by the preinjection of CaN inhibitors. Cofilin activation could be further demonstrated by increased actin–cofilin binding and a significant depolymerization of neuronal actin, both of which were also blocked by CaN inhibitors. Finally, we demonstrated a CaN-dependent loss of dendritic spines histologically. DISCUSSION: The data demonstrate a CaN-dependent, cellular mechanism through which prolonged seizure activity results in loss of dendritic spines via cofilin activation. Further research into this area may provide useful insights into the pathology of SE and epileptogenic mechanisms.

Food Restriction Alters Neuronal Morphology in the Hypothalamic Ventromedial Nucleus of Male Rats

Several lines of evidence have implicated the hypothalamic ventromedial nucleus (VMH) in the control of caloric homeostasis. For example, the activity of VMH neurons depends on energy availability. We tested the hypothesis that energy balance may involve the remodeling of the dendritic arbor of VMH neurons. We compared two groups of animals: one group had ad libitum access to food, and the other experienced 10-d restricted access to food. As expected, the food-deprived group lost body weight and had reduced levels of glucose, insulin, and leptin. VMH neurons were visualized after Golgi impregnation, and dendrite length was measured. Food deprivation had differential effects on VMH neurons. In particular, within the ventrolateral VMH, for neurons with long primary dendrites (LPDs) that extended in the lateral, but not medial, direction, the LPDs were 31% shorter. These same neurons exhibited a 32% reduction in the number of other dendrites without a change in soma size. In contrast, within the dorsomedial VMH, for neurons with medially, but not laterally, extended LPDs, the soma area was reduced by 28%. However, neurons in the dorsomedial VMH did not display a change in the length or number of dendrites, regardless of LPD direction. Thus, although structural changes during calorie depletion occur in both the dorsomedial and ventrolateral VMH, only the latter exhibits a remodeled dendritic arbor. These results also suggest that the direction of the LPD may be an important marker of neuronal function in the VMH.